This Phase II Advanced Technology SBIR [PA-06-134] proposal is in response to NIAID's Notice of High-Priority Influenza Research Areas [NOT-AI-05-013]. In order to provide health officials with the tools required to efficiently combat a pandemic strain of influenza, it is essential that rapid and cost-effective methods for vaccine production be developed. The proposed instrument is specifically designed to provide a rapid count of the number of "intact" viruses per unit volume in liquid. While optimization of vaccine production methods is an example application for which the number of intact viruses is a valuable measurement, the time and cost savings of virus enumeration within minutes, rather than days, is anticipated to have substantial impact on several other virus-related fields. Support for that outlook is provided by the excellent group of collaborators we have been able to assemble during the Phase I efforts, including scientists from two large vaccine manufacturers (MedImmune and Novartis), a small cutting-edge vaccine manufacturer (Protein Sciences Corporation), a biological reagents company (Microbix Biosystems, Inc.), a diagnostics company (Quidel Corporation), and a College of Medicine (at Baylor). All Phase I objectives were achieved and in many cases exceeded. During Phase II efforts we will i) design and construct an advanced prototype Virus CounterTM and develop automated methods for instrument parameter setup; ii) evaluate instrument performance in comparison with standard methods such as plaque assays, fluorescence focus assays, transmission electron microscopy and real-time quantitative PCR, iii) optimize the Virus CounterTM and assay for utility in vaccine production, and iv) conduct beta-site testing of the instrument. [unreadable] [unreadable] PUBLIC HEALTH RELEVANCE: In order to provide health officials with the tools required to efficiently combat a pandemic strain of influenza, it is essential that rapid and cost-effective methods for vaccine production be developed. The proposed instrument is specifically designed to provide a rapid count of the number of "intact" viruses per unit volume in liquid. While optimization of vaccine production methods is an example application for which the number of intact viruses is a valuable measurement, the time and cost savings of virus enumeration within minutes, rather than days, is anticipated to have substantial impact on several other virus-related fields. [unreadable] [unreadable] [unreadable]